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PubMed2004

Egyptian genotyping of Giardia lamblia.

Out of 105 patients infected with Giardia, 38 patients have Genotype I (36.19%), 13 have Genotype II (12.38%), 10 have Genotype III (9.52%), 16 have mixed Genotype infection (15.24%) and 28 with undetermined Giardia infection by PCR (26.67%). None of the control group gave positive results for Giardia in stool by PCR. So, the sensitivity of the test for detection and identification of Giardia Genotypes from the original stool samples was 73.33% and specificity was 100%. Out of 61 cases in the symptomatic group, the prevalence of Giardia Genotype I was 32.79%, Genotype II was 16.39%, Genotype III was 9.84%, mixed Genotype infection was 16.39% and undetermined Genotype was 24.59% as compared to 40.91%, 6.82%, 9.09%, 13.64% & 29.55% in the asymptomatic group respectively. There is statistically insignificant difference between both groups as regarding the prevalence of the different Giardia Genotypes. (P < or = 0.05). The use of PCR as a routine work for diagnosis of giardiasis is not accepted at least in the developing and under-developing countries due to its high cost, the high quality of technical staff and advanced laboratory equipments required for PCR performance. Its application is usually limited to research activities, the detection of water sources contamination and for the detection of a potential source of Giardia infection in epidemics.

Adolescent↗

PubMed2005

Contribution of cattle farms towards river contamination with Giardia cysts and Cryptosporidium oocysts in Sungai Langat Basin.

A study to determine the contribution of Giardia cysts and Cryptosporidium oocysts from cattle farms was carried out at the Langat Basin. This study investigated the contribution of cattle farms, located near Sungai Langat and Sungai Semenyih, towards river contamination with these cysts and oocysts. The findings showed that out of 24 samples of water taken from Sungai Semenyih, 4.2% was positive for Giardia cysts with a concentration of 1.3 cysts/L and 20.8% were positive with Cryptosporidium oocysts with a range of 0.7 - 2.7 oocysts/L. At Sungai Langat, from the 43 samples taken, 23.3% were positive for Giardia cysts with a range of 1.5 - 9 cysts/L whereas 11.6% were positive with Cryptosporidium oocysts with a range of 2.5 - 240 oocysts/L. Isolation of cysts and oocysts in bovine faecal materials revealed that 14.6% of faecal samples were positive for Giardia cysts which had a range of 75 - 1.3x104 cysts/g and 25% were positive for Cryptosporidium oocysts with a range of 50 - 3.9x105 oocysts/g. From the cattle wastewater, 98% were positive with oocysts and 6.7% with cysts. The concentrations were between 20 - 3.1x103 oocysts/mL for Cryptosporidium and 4 - 75 cysts/mL for Giardia. Given that the prevalence of Cryptosporidium and Giardia are high amongst the cattle and the positive findings of the (oo)cysts in the river samples, it could be deduced that there is a very high possibility of the cattle farms contaminating the river with Giardia cysts and Cryptosporidium oocysts. Viability study of Cryptosporidium oocysts in the surrounding soil and pond within the cattle farm showed that the viability of Cryptosporidium oocysts decreased with time. It was estimated that it will take 52 days for all the oocysts from both environment to be non-viable. With a viability rate of approximately 2 months in a cattle farm setup, river water contaminated with Cryptosporidium oocysts has a high chance of acting as an agent of transmission. As cattle farms are also inhabited by the owners and their families, this problem may pose a threat to humans (e.g. children) especially if they are dependent on the river water as their source of water for their daily activities.

Animal Husbandry↗

PubMed1989

Antigenic conservation and variation in Giardia cysts from various vertebrate hosts.

Monoclonal antibodies produced against Giardia muris cysts reacted in indirect immunofluorescence with homologous cysts and cysts from a Giardia-infected wild Norway rat but did not cross-react with Giardia lamblia cysts of human, dog, or beaver sources. Another monoclonal antibody raised against Giardia simoni cysts from the Norway rat reacted with homologous cysts (rat) and cross-reacted with cysts from a cow. The demonstration of antigenic differences at the cyst surfaces of Giardia organisms of animal and human origin suggests that it is possible to identify the animal source of Giardia cysts according to their pattern of reactivity with monoclonal antibodies. Such identification would have a useful application in affirming the possible zoonotic transmission of animal source Giardia species to humans.

Animals↗

PubMed1988

A comparison of Giardia microti and Spironucleus muris cysts in the vole: an immunocytochemical, light, and electron microscopic study.

We have shown that cysts of the genus Spironucleus share many common morphological features with Giardia cysts including: 2-4 nuclei, flagellar axonemes, a distinct cyst wall, and they even display the same immunostaining as Giardia cysts when labeled with antibodies specific for Giardia cyst wall. A direct comparison of Spironucleus muris and Giardia microti cysts have revealed that cysts of S. muris are significantly smaller than cysts of G. miroti. At the ultrastructural level, the cyst walls are similar in fibrillar appearance, but the width of the S. muris cyst wall is significantly less than that of G. microti. The cysts of S. muris also differ from G. microti in that they contain a striated rootlet fiber, flagellar sheath, and numerous glycogen rosettes. Characteristic features of Giardia include the adhesive disc and median body. Although the cysts of Spironucleus and Giardia are similar in appearance, these unique morphological features can be used to distinguish between the 2 protozoa and should be employed in the detection of Giardia cysts in water samples.

Animals↗

PubMed1987

SEM evidence for a new species, Giardia psittaci.

The genus Giardia has been subdivided by Filice (1952) into 3 species, G. agilis, G. muris, and G. duodenalis, based on the morphology of the median body and subtle variations in the dimensions of trophozoites. Giardia trophozoites were isolated from the small intestine of budgerigars (parakeets) and examined morphologically with light and scanning electron microscopy. These trophozoites, like other Giardia spp., possessed a flattened dorso-ventral shape, 8 flagella, and an adhesive disc on the ventral surface. The presence of a claw hammer-shaped median body suggested classification of these trophozoites as G. duodenalis. However, unlike any known members of G. duodenalis, the Giardia trophozoites from budgerigars were morphologically distinct in that they lacked the ventrolateral flange and therefore did not have a marginal groove bordering the anterior and lateral border of the adhesive disc. This distinct morphology clearly indicated that trophozoites from budgerigars should be considered as a separate species, G. psittaci. Our evidence has demonstrated that median body shape cannot serve as a sole criterion for speciation of Giardia. In addition, if other avian species of Giardia also resemble G. psittaci, then this would suggest that evolutionary divergence has occurred in the genus Giardia.

Adhesiveness↗

PubMed1986

Lack of cellular cytotoxicity by human mononuclear cells to Giardia.

Cytotoxicity of mononuclear leukocytes (MNL) to Giardia lamblia was compared by using two techniques. The first method assessed the viability of surviving Giardia directly by culturing and the second method measured release of incorporated [3H] thymidine. Cytotoxicity, as measured directly by culturing and visual assessment, showed that the numbers of surviving Giardia decreased over time whether cultured with or without MNL but that Giardia survived significantly better in the presence of MNL at 18 hr (21.8 +/- 8.3% with MNL compared with 3.4 +/- 1.5% without MNL). Release of [3H]thymidine increased whether Giardia were cultured with or without MNL and although there was a tendency for increased release with MNL, there was no significant difference. Dead labeled Giardia released significantly more label in the presence of MNL than without MNL, suggesting that MNL cause release of [3H]thymidine after phagocytosis. The thymidine release assay therefore does not measure spontaneous cytotoxicity of MNL to Giardia.

Animals↗

PubMed1994

GP49, an invariant GPI-anchored antigen of Giardia lamblia.

Giardia lamblia is a primitive protozoan and a major cause of waterborne enteric disease throughout tropical and temperate zones. The ability to grow the infective trophozoites in culture as well as the discovery of the method of in vitro encystation made it possible to study the biology of this primitive protozoan and to characterize the surface antigens. Giardia trophozoites are exposed to high concentrations of fatty acids in the human small intestine. This raises the possibility that intestinal fatty acids may become incorporated into Giardia. Therefore, we determined the pattern of fatty acylation of Giardia surface molecules. By metabolic labeling with radiolabeled fatty acids we identified a single glycosylphosphatidylinositol (GPI)-anchored surface protein in Giardia. GP49 differs from the cysteine-rich variable surface antigens described previously. The presence of a GPI anchor in GP49 was supported by the metabolic incorporation of [14C]-ethanolamine, [3H]-myoinositol and fatty acids into the protein. This was confirmed by chemical and enzymatic cleavage experiments. Most interestingly, GP49 was found to be present in different isolates of Giardia and thus can be considered as an invariant surface antigen. Although the biological function of GP49 is not known, recently we have found that intact and soluble GP49 altered the electrolyte fluxes which regulate fluid secretion in the cultured human intestinal epithelial cell line, T84. These studies indicate that the GPI-anchored invariant antigen of Giardia may play an important role in the pathophysiology of giardiasis.

Animals↗

PubMed1996

The efficacy of a Giardia lamblia vaccine in kittens.

Twenty kittens were vaccinated with a Giardia lamblia vaccine prepared on a commercial scale on day 0 and boosted on day 21 (group 1); while 10 kittens received only saline (group 2). These kittens were challenged on day 35 with 10(6) Giardia lamblia trophozoites by a surgical intraduodenal injection. Three control kittens were not vaccinated and not challenged (group 3). Following challenge, Giardia vaccinated kittens had significantly fewer days in which abnormal stools were observed and reduced food intake occurred compared to saline injected animals. The rate of weight gain between group 1 and group 2 animals was not different in the prechallenge period (day 0 to day 35), but vaccinated animals had a significantly higher weight gain in the postchallenge period (P < 0.05). On day 56, all vaccinated animals were not passing cysts in their feces, while 40% of saline injected kittens had Giardia cysts in their feces. In vaccinated kittens, cysts were never demonstrated in 45% of the animals, while cysts were detected in 90% of the saline injected kittens. Viability of the cysts in vaccinated kittens was 38% while the cysts viability in saline injected kittens was 99%. On postmortem examination, trophozoites could be detected in 5% of vaccinated kittens and 60% of saline injected kittens. Vaccination produced an elevated Giardia specific serum IgG and IgA response prior to challenge and throughout the postinfection period. The Giardia infection in the saline injected group did not induce an elevated specific serum response. Giardia vaccination of kittens provides protection in kittens from an experimental challenge by reducing or eliminating intestinal trophozoites and fecal cyst excretion.

Aging↗

PubMed1991

Giardia lamblia and Cryptosporidium infections in child day-care centers in Fulton County, Georgia.

Risk factors for the introduction, spread and persistence of Cryptosporidium and Giardia lamblia infections in child day-care centers are not well understood. In 1989 and 1990 stool specimens were obtained from 292 diapered children attending 17 randomly selected day-care centers in Fulton County, GA; 8 (2.7%) children in 2 centers were infected with Cryptosporidium and 21 (7.2%) children in 7 centers were infected with Giardia. In 1986 the prevalence of Cryptosporidium and Giardia in these same centers had been 0.4 and 11.0%, respectively; the prevalence of Cryptosporidium, but not Giardia, increased significantly (P = 0.04) between 1986 and 1989 to 1990. Risk factors for Giardia infection included day-care attendance for greater than 3 months, the presence of toddlers in the classroom and the presence of other children in the household. Day-care centers with a Giardia-positive child in 1986 were not more likely to have an infected child in 1989 to 1990. Cryptosporidium, like Giardia, may be endemic in day-care centers in Fulton County.

Child Day Care Centers↗

PubMed1990

Prevalence of Giardia spp. in beaver and muskrat populations in northeastern states and Minnesota: detection of intestinal trophozoites at necropsy provides greater sensitivity than detection of cysts in fecal samples.

Surveys of the prevalence of the intestinal protozoan Giardia spp. in animal populations have relied almost exclusively on the detection of cysts in fecal samples. We have determined the prevalence of Giardia spp. in beaver and muskrat populations in four northeastern states and Minnesota by using both the detection of trophozoites in mucosal scrapings from live-trapped animals at necropsy and the detection of cysts in fecal samples collected from kill-trapped animals. In muskrats the prevalence of Giardia infection was 36.6% by cyst detection in fecal samples (n = 790) from kill-trapped animals and 95.9% in live-trapped muskrats when the intestinal contents were analyzed for the presence of trophozoites (n = 219). Similarly, in beavers, Giardia infection was 9.2% by cyst detection in fecal samples (n = 662) from kill-trapped beavers and 13.7% in live-trapped animals examined for the presence of intestinal trophozoites (n = 302). The detection of trophozoites in mucosal scrapings from live-trapped animals consistently yielded a significantly higher prevalence for both muskrats and beavers than did the method based on detection of cysts in the fecal samples. The prevalence of Giardia infection in juvenile and adult live-trapped muskrats was similar (92.5 and 94.4%, respectively), but the prevalence in juvenile live-trapped beavers (23.2%) was significantly greater than that seen in the adult animals (12.6%). No difference in Giardia prevalence on the basis of sex was seen in either animal species. Regional variation, often statistically significant, was seen in the prevalence of Giardia in beavers in the northeastern states and Minnesota, but was not detected for muskrats.

Animals↗

PubMed1991

Controlled trial of Giardia lamblia: control strategies in day care centers.

BACKGROUND: Strategies for control of Giardia lamblia in day care differ in numbers of children treated and in costs to parents and day care operators. The effectiveness of these strategies has not been systematically evaluated. METHODS: We conducted a prospective randomized controlled trial comparing three strategies for control of Giardia in infant-toddler day care centers: Group 1, exclusion and treatment of symptomatic and asymptomatic infected children; Group 2, exclusion and treatment of symptomatic infection only; Group 3, exclusion and treatment of symptomatic infection, treatment of asymptomatic infection in the center. The study included 31 day care centers with 4180 child-months of observation. Giardia prevalence was determined before intervention and 1, 2, 4, and 6 months later; new infants and toddlers were tested on admission. RESULTS: Initial Giardia prevalences were 18% to 22% in the three groups. Giardia was identified in 10.5% of 676 new infants and toddlers entering study day care centers during the 6-month follow-up. Giardia prevalences by intervention group were 8%, 12%, and 7% at 1 month, and 7%, 8%, and 8% at 6 months. CONCLUSIONS: The stricter intervention resulted in greater cost in terms of child day care and parents' work days lost, but did not result in significantly better control of Giardia infections in this day care environment.

Child Day Care Centers↗

PubMed1989

[The frequency and geographic distribution of Giardia infections in ruminants in Switzerland].

In various districts (Cantons) of Switzerland 815 calves (a few days up to 6 months old), 382 lambs and 20 young goats (both groups 1-6 months old) were selected randomly for a single coprological examination (flotation method) for Giardia infection. In average 26.6% of the calves, 29.8% of the lambs and 4 of 20 young goats excreted Giardia cysts. In 9 Cantons the percentages of cyst excretors among the calves varied between 15 and 32, but these differences were not significant. Further there were no significant differences in the frequency of cyst excretion between calves of 3 different breeds and 2 age groups (up to 3 months old and 3 to 6 months). The intensity of cyst excretion was high and varied in random samples between 4.1 x 10(3) and 3.0 x 10(5) cysts per g of faeces in calves and between 2.2 x 10(3) and 1.6 x 10(5) in lambs. In 5 of 7 farms where calves and lambs were maintained simultaneously both animal species were Giardia infected. As expected, trophozoites of Giardia isolates from calves and lambs belonged to the Giardia duodenalis type and could not be differentiated on a morphological basis. Giardia cysts from calves (average measurements: 13.7 x 9.1 microns) and lambs (13.8 x 9.2 microns) were indistinguishable both morphologically and morphometrically. The results indicate that Giardia infections are frequent and geographically widely distributed in calves and lambs in Switzerland.

Animals↗

PubMed1997

Giardia and Cryptosporidium in dairy calves in British Columbia.

A study was undertaken to determine the prevalence of Giardia infections in dairy calves and to compare Giardia and Cryptosporidium infections in calves of different ages. Fresh fecal samples were collected from 386 male and female Holstein calves (newborn to 24 wk) in 20 dairies located in the lower Fraser river valley area of British Columbia. Giardia intestinalis, Cryptosporidium parvum, and Cryptosporidium muris were enumerated in each sample after concentration by sucrose gradient centrifugation and immunofluorescent staining. Giardia was identified at all farm locations. The overall prevalence of Giardia in calves was 73% with a geometric mean cyst count of 1180 cysts per gram of feces (CI, 41 to 5014). Cryptosporidium parvum and C. muris were identified in 80% and 40% of the farms, respectively. The prevalence of C. parvum was 59%, and the geometric mean for oocysts was 457 oocysts per gram of feces (CI, 18 to 160). The prevalence of C. muris was only 2% and the mean oocyst counts were 54 oocysts per gram of feces. Giardiasis was not age dependent, and approximately 80% of the calves from 2 to 24 wk were infected. In contrast, C. parvum infections were predominant in calves 2 to 4 wk, while C. muris was demonstrated in calves older than 4 wk. Fourty-seven percent of calves with diarrhea had high numbers of Giardia cysts in their feces. Giardia infections are highly prevalent in dairy calves and should be considered in animals with diarrhea or failure to thrive.

Animals↗

PubMed2005

Development of species-specific rDNA probes for Giardia by multiple fluorescent in situ hybridization combined with immunocytochemical identification of cyst wall antigens.

In this study, we describe the development of fluorescent oligonucleotide probes to variable regions in the small subunit of 16S rRNA in three distinct Giardia species. Sense and antisense probes (17-22 mer) to variable regions 1, 3, and 8 were labeled with digoxygenin or selected fluorochomes (FluorX, Cy3, or Cy5). Optimal results were obtained with fluorochome-labeled oligonucleotides for detection of rRNA in Giardia cysts. Specificity of fluorescent in situ hybridization (FISH) was shown using RNase digestion and high stringency to diminish the hybridization signal, and oligonucleotide probes for rRNA in Giardia lamblia, Giardia muris, and Giardia ardeae were shown to specifically stain rRNA only within cysts or trophozoites of those species. The fluorescent oligonucleotide specific for rRNA in human isolates of Giardia was positive for ten different strains. A method for simultaneous FISH detection of cysts using fluorescent antibody (genotype marker) and two oligonucleotide probes (species marker) permitted visualization of G. lamblia and G. muris cysts in the same preparation. Testing of an environmental water sample revealed the presence of FISH-positive G. lamblia cysts with a specific rDNA probe for rRNA, while negative cysts were presumed to be of animal or bird origin.

Animals↗

PubMed2000

PCR detection of Giardia lamblia in stool: targeting intergenic spacer region of multicopy rRNA gene.

A PCR based detection that amplifies the 552-bp intergenic spacer (IGS) region of multicopy rRNA gene of Giardia lamblia and 320-bp internal sequences to first PCR product has been used in diagnosis of giardiasis in stool sample. The primers were found highly specific to Giardia spp. only, because no amplification was observed with DNAs from other enteric pathogens like Escherichia coli, Shigella dysenteriae and Entamoeba histolytica. The test could detect even less than 2 pg of genomic DNA from Giardia trophozoites. In direct diagnosis of Giardia lamblia in stool samples, it was observed that PCR amplification of IGS followed by nested PCR could enhance the sensitivity and specificity of the tests manifold and the system was able to detect as low as 10 parasites in 100 microl of stool. The comparative evaluation of the present system with conventional microscopy, CIEP and ELISA in the diagnosis of giardiasis from diarrhoeic stool samples and control subjects demonstrated a 100% correlation among nested PCR, microscopic examination and ELISA in patients suggestive of giardiasis (Group I) and control subjects (Group II). In Group I cases (patients suffering from other than giardiasis), CIEP, ELISA and nested PCR showed better results than microscopic examination. However, among them, PCR was found most sensitive and specific because 20% positivity was noticed by PCR whereas CIEP and ELISA showed only 7.14% and 12.85%, respectively. Break-up results showed that all the samples which were positive by CIEP or ELISA, also found positive by PCR. The present observation clearly suggests the use of PCR that amplifies the intergenic spacer region of multicopy rRNA gene of Giardia lamblia followed by nested PCR for routine, quick and reliable detection of Giardia lamblia in stool samples.

Animals↗

PubMed1980

Axenic growth of Giardia lamblia in Diamond's TPS-1 medium.

A strain of Giardia lamblia was gradually adapted to grow in Diamond's TPS-1 medium. The parasites grew luxuriantly and formed a monolayer on the surface of the culture vessels. When G. lamblia was inoculated into fresh medium, the population of the parasites grew from 22,000 cells/ml to peak numbers of 2.1 X 10(6)/ml in six days with a generation time of 12.2 hours at 37 degrees C. After a few days of incubation a sediment could be seen at the bottom of the tubes which contained a multitude of free-swimming Giardia as well as clumps of them. The sediment sometimes also contained many rings of Giardia. The rings were usually associated with dead Giardia cells containing a large bubble. Two, three and sometimes as many as six Giardia were seen surrounding the bubble. The bubble probably provides additional surface area to which the Giardia may attach.

Animals↗

PubMed1996

Characterisation and purification of pyruvate:ferredoxin oxidoreductase from Giardia duodenalis.

The major 2-oxoacid oxidoreductase (2-OR), pyruvate:ferredoxin oxidoreductase (PFOR) from Giardia duodenalis has been purified to apparent homogeneity. A second 2-OR with a preference for alpha-ketobutyrate as substrate was identified and was removed from PFOR containing fractions during purification. Only PFOR and the second 2-OR were identified in gels of crude Giardia extracts assayed for 2-OR activity. The native form of PFOR which is membrane associated, is a homodimer of 138 kDa subunits. Pyruvate is the preferred substrate: alpha-ketobutyrate and oxaloacetate, but not phenyl-pyruvate or alpha-ketoglutarate, are decarboxylated. PFOR from Giardia is more stable than PFOR from most other organisms and purified PFOR can be stored without deterioration at -70 degrees C. Purified PFOR donates electrons to Giardia ferredoxin (Fd I) with concomitant reduction of metronidazole. However, two other Giardia ferredoxins did not accept electrons from PFOR. Consistent with the involvement of PFOR in metronidazole activation, the activity of pyruvate dependent 2-OR activity was decreased in all metronidazole-resistant lines tested but not in furazolidone-resistant lines. The presence of three different ferredoxins and two 2-ORs in Giardia suggests that a number of different electron transport pathways operate in this organism providing unusual metabolic flexibility for a eukaryote.

Animals↗

PubMed1990

Axenic isolation of Giardia strains from primates and rodents.

During the examination of animals at the Poznan Zoological Gardens, attempts were made to isolate Giardia strains. Using an in vitro excystation procedure, eight samples of cysts from animals with asymptomatic giardiasis were inoculated on BI-S-33 medium. The ease of isolation and axenization of Giardia was surprising; five axenic isolates of Giardia, belonging to the G. duodenalis morphological group, were established from primates (slow loris, lesser slow loris and siamang) and from rodents (Gambian giant pouched rat and cuis). The growth of all isolates was abundant and similar; the peak number of trophozoites on the seventh day (depending on the Giardia isolate) was 2.3 X 10(6)-3.2 X 10(6) and generation times were 8.2-19.3 h. The easy establishment of these isolates confirmed that they belong to the G. duodenalis morphological group. The recent hypothesis that Giardia may be introduced to a human population from an animal source implies the necessity to isolate and differentiate parasite strains from various hosts. In this respect, the first isolation of Giardia strains from non-human primates and from rodents is of particular importance.

Animals↗